Determining potential immunomodulatory drug efficacy in sepsis using ELISpot

Sci Rep. 2025 Apr 18;15(1):13464. doi: 10.1038/s41598-025-92016-6.

Abstract

This study evaluated the ability of ELISpot to identify potential immuno-modulatory drug therapies in sepsis. ELISpot was performed ex vivo on whole blood from septic patients and healthy controls. Innate and adaptive immunity were evaluated by production of TNF-α and IFN-γ, respectively. Drug efficacy was determined by their effects to modulate the both the number of cytokine-producing cells and amount of cytokine produced per cell. The corticosteroid dexamethasone was evaluated for its ability to down modulate TNF-α and IFN-γ production. The TLR7/8 agonist resiquimod (R848) and T cell stimulants IL-7 and anti-PD-1 mAb were tested for their ability to enhance immunity. LPS and resiquimod increased total TNF-α production in septic patients by 1,549% and 1,829%, respectively. Conversely, dexamethasone diminished the responses to LPS or resiquimod by 75% and 61%, respectively. IL-7, but not anti-PD-1 mAb markedly increased IFN-γ production in both healthy subjects (121%) and septic patients (82%). Dexamethasone also reduced anti-CD3/CD28 mAb stimulated IFN-γ production by 69%; while IL-7 ameliorated dexamethasone-induced suppression. IL-7 significantly enhanced lymphocyte function in over 90% of septic patients. ELISpot can reveal host immune response patterns and the effects of drugs to selectively down- or up-regulate patient immunity. Furthermore, the ability of ELISpot to detect the effect of specific immuno-modulatory drugs to independently regulate the innate and adaptive host response could enable precision-based immune drug therapies in sepsis.

Keywords: Adaptive immunity; Anti-PD-1; Checkpoint inhibitors; Corticosteroids; IL-7; Innate immunity.

MeSH terms

  • Adaptive Immunity / drug effects
  • Adult
  • Aged
  • Cytokines
  • Dexamethasone / pharmacology
  • Dexamethasone / therapeutic use
  • Enzyme-Linked Immunospot Assay* / methods
  • Female
  • Humans
  • Imidazoles / pharmacology
  • Immunity, Innate / drug effects
  • Immunologic Factors* / pharmacology
  • Immunologic Factors* / therapeutic use
  • Immunomodulating Agents* / pharmacology
  • Immunomodulating Agents* / therapeutic use
  • Interferon-gamma / metabolism
  • Interleukin-7 / pharmacology
  • Lipopolysaccharides / pharmacology
  • Male
  • Middle Aged
  • Sepsis* / drug therapy
  • Sepsis* / immunology
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Dexamethasone
  • resiquimod
  • Interferon-gamma
  • Imidazoles
  • Tumor Necrosis Factor-alpha
  • Lipopolysaccharides
  • Immunomodulating Agents
  • Interleukin-7
  • Cytokines
  • Immunologic Factors